The Effect of Neutropenia and Filgrastim (G-CSF) in Cancer Patients With COVID-19 Infection.
Neutropenia is commonly encountered in cancer patients, and recombinant human granulocyte colony-stimulating factor (G-CSF, filgrastim) is widely given to oncology patients to counteract neutropenia and prevent infection. G-CSF is both a growth factor and cytokine that initiates proliferation and differentiation of mature granulocytes. However, the clinical impact of neutropenia and G-CSF use in cancer patients, who are also afflicted with coronavirus disease 2019 (COVID-19), remains unknown. An observational cohort of 304 hospitalized patients with COVID-19 at Memorial Sloan Kettering Cancer Center was assembled to investigate links between concurrent neutropenia (N=55) and G-CSF administration (N=16) on COVID-19-associated respiratory failure and death. These factors were assessed as time-dependent predictors using an extended Cox model, controlling for age and underlying cancer diagnosis. To determine whether the degree of granulocyte response to G-CSF affected outcomes, a similar model was constructed with patients that received G-CSF, categorized into high- and low-response, based on the level of absolute neutrophil count (ANC) rise 24 hours after growth factor administration. Neutropenia (ANC < 1 K/mcL) during COVID-19 course was not independently associated with severe respiratory failure or death (HR: 0.71, 95% Cl: 0.34-1.50, P value: 0.367) in hospitalized COVID-19 patients. When controlling for neutropenia, G-CSF administration was associated with increased need for high oxygen supplementation and death (HR: 2.97, 95% CI: 1.06-8.28, P value: 0.038). This effect was predominantly seen in patients that exhibited a high response to G-CSF based on their ANC increase post-G-CSF administration (HR: 5.18, 95% CI: 1.61-16.64, P value: 0.006). Possible risks versus benefits of G-CSF administration should be weighed in neutropenic cancer patients with COVID-19 infection, as G-CSF may lead to worsening clinical and respiratory status in this setting.


Fourier interpolation stochastic optical fluctuation imaging.
Stochastic Optical Fluctuation Imaging (SOFI) is a super-resolution fluorescence microscopy technique which allows to enhance the spatial resolution of an image by evaluating the temporal fluctuations of blinking fluorescent emitters. SOFI is not based on the identification and localization of single molecules such as in the widely used Photoactivation Localization Microsopy (PALM) or Stochastic Optical Reconstruction Microscopy (STORM), but computes a superresolved image via temporal cumulants from a recorded movie. A technical challenge hereby is that, when directly applying the SOFI algorithm to a movie of raw images, the pixel size of the final SOFI image is the same as that of the original images, which becomes problematic when the final SOFI resolution is much smaller than this value. In the past, sophisticated cross-correlation schemes have been used for tackling this problem. Here, we present an alternative, exact, straightforward, and simple solution using an interpolation scheme based on Fourier transforms. We exemplify the method on simulated and experimental data.


Treatment of posttransplant lymphoproliferative disease in the central nervous system of a lung transplant recipient using allogeneic leukocytes.
Posttransplant Epstein-Barr virus-related lymphoproliferative disease (PT-LPD) is a common and often fatal complication following solid organ and hematopoietic stem cell transplantation. PT-LPD following solid organ transplantation generally occurs in B cells of recipient origin in contrast to PT-LPD in marrow transplant recipients, which is exclusively of donor origin. The efficacy of adoptive immunotherapy using donor leukocytes to treat PT-LPD in bone marrow transplant recipients has recently been reported. Because PT-LPD in solid organ transplant recipients is generally of recipient origin, the potential application of adoptive immunotherapy of PT-LPD in solid organ recipients obligates the use of either autologous or allogeneic HLA identical leukocytes, with the attendant risk of organ rejection if cells mismatched with the transplanted organ are used. Nonirradiated allogeneic mononuclear cells from an Epstein-Barr virus (EBV)-seropositive, HLA-identical normal sibling were used to treat a monoclonal EBV lymphoma of recipient origin in the central nervous system of a child who had undergone an HLA-mismatched cadaveric lung transplant. The patient received three separate mononuclear cell infusions over a 9-month period, each containing 1 x 10(6) CD3+ mononuclear cells per kilogram. Complete clinical, radiological, and pathological remission was achieved with this treatment regimen. The response correlated with in vivo reconstitution of normal EBV-specific cytotoxic activity and cytotoxic T lymphocyte precursor frequency. Use of allogeneic HLA-compatible mononuclear cells may thus offer an additional mode of therapy for EBV-related lymphoproliferative disease in selected solid organ transplant recipients refractory to conventional therapies.


Gender, age, and body weight are the major predictive factors for bone mineral density in Crohn's disease: a case-control cross-sectional study of 113 patients.
We conducted this study to assess bone mineral density and to evaluate conceivable predictive factors for bone loss in patients with Crohn's disease. One hundred-thirteen patients with Crohn's disease and 113 healthy subjects, individually matched for gender, age, and body weight were investigated. The group consisted of 68 women and 45 men. The median duration of Crohn's disease was 6 yr. Two-thirds of the patients had been subjected to intestinal resection. Seventy-seven percent had at some time been treated with corticosteroids. Bone mineral density in the lumbar spine, the hip, and the total body skeleton was measured by dual-energy X-ray absorptiometry (DEXA). In patients with Crohn's disease bone mineral density was not different from that of healthy controls except for a regional decrease in bone mineral density of the hip in female patients. The strongest predictors of bone mineral density were gender, age, and body weight. Corticosteroid use was only a weak predictor of diminished bone density. Duration of disease and intestinal resection had no predictive value for bone mineral density. Gender, age, and body weight are the major determinants of bone mineral density in patients with Crohn's disease. As in healthy individuals, the combined effect of these factors account for up to 50% of the variability in bone mineral density.


Identification of gallbladder mucin-bilirubin complex in human cholesterol gallstone matrix. Effects of reducing agents on in vitro dissolution of matrix and intact gallstones.
The goals of this study were to isolate and characterize the nonlipid matrix of human cholesterol gallstones. The lipid portion of gallstones was dissolved in ethanol/ether, leaving an insoluble, granular, brown-black matrix that constituted 12.5% of solitary large stones and 3.5% of multiple small stones. The matrix was partially solubilized by sonication and studied by exclusion gel chromatography and density gradient ultracentrifugation. On Sepharose 2B column chromatography, bile pigment eluted with glycoprotein in the void volume, suggesting the presence of a high molecular weight complex (Mr greater than 2 X 10(6)). The identity of mucin in this complex was confirmed by its typical buoyant density during ultracentrifugation. The major bile pigments in the matrix were identified as bilirubin (84%) and bilirubin monoglucuronide (15%) by thin-layer chromatography. Because of their ability to solubilize mucin-type glycoproteins, we tested the ability of the reducing agents 2-mercaptoethanol (2ME) and N-acetylcysteine (NAcCys) to solubilize gallstone matrix. Both reducing agents caused a two- to threefold enhancement of matrix dissolution after 4 d compared to aqueous buffer alone (P less than 0.01). Sepharose 2B chromatography revealed that 2ME released a high molecular weight mucin-bilirubin complex as well as unbound pigment from the insoluble matrix. We also tested the effect of reducing agents on dissolution of matched cholesterol gallstones by monooctanoin, a cholesterol solvent. Both 2ME and NAcCys significantly accelerated gallstone dissolution in monooctanoin. Matched human cholesterol stones (n = 10) incubated for 4 d in monooctanoin plus either 2ME or NAcCys (1 M final concentration) weighed approximately half as much (P less than 0.01 for each) as stones incubated in monooctanoin alone. This study describes, for the first time, the isolation of a bilirubin-mucin complex in the insoluble matrix of human cholesterol gallstones. The ability of reducing agents to dissolve the matrix and thereby accelerate gallstone dissolution by monooctanoin in vitro may be relevant to gallstone dissolution in humans.


Emerging role of embryo secretome in the paracrine communication at the implantation site: a proof of concept.
To assess the role of embryo secretome in modifying the molecular profile of glycodelin A (GdA) in endometrial organoids (ORG) mimicking the implantation window. To verify whether the use of embryo-conditioned culture medium at the time of the embryo transfer may increase in vitro fertilization outcome. Molecular study with human endometrial ORG and embryo-conditioned culture medium. Retrospective study using prospectively recorded data. University hospital. For isolation and culture of endometrial glandular ORG, endometrial biopsy specimens from five white women of proven fertility undergoing laparoscopy for tubal sterilization. A total of 75 women undergoing intracytoplasmic sperm injection for tubal and/or male infertility factor. In vitro fertilization. Pinopodes presence in human endometrial ORG. Glycodelin A expression profile by means of two-dimensional electrophoresis. In vitro fertilization outcome. This in vitro study demonstrated that the treatment of endometrial ORG with the secretome of medium conditioned by the growing embryo increased the GdA relative abundance and induced a different glycoform pattern. Biochemical and clinical pregnancy rate significantly increased when the spent medium was loaded during the transfer (17.5% vs. 36.6% and 16.5% vs. 35.1%, respectively). This study demonstrated that the secretome of implanting embryos is able to induce the expression as well as to determine the relative abundance and the glycosilation profile of endometrial GdA, a protein having a key role in the embryo-endometrial cross talk. Moreover, a significant increase in pregnancy rate was observed when the embryo transfer was performed by using the culture medium conditioned by the growing embryo.


Polydatin inhibits cell proliferation, invasion and migration, and induces cell apoptosis in hepatocellular carcinoma.
Polydatin, a small molecule from Polygonum cuspidatum, has many biological functions, particularly anti-cancer effects. However, the anti-cancer effects of polydatin in hepatocellular carcinoma (HCC) have not been examined yet. In the present study, MTT assay, BrdU assay, transwell invasion assay, and wound healing assay were performed to determine cell proliferation, invasion and migration. Flow cytometry and TUNEL assay were used to measure cell apoptosis. Quantitative real-time PCR and western blotting assays were used to determine mRNA and protein expression levels. Xenograft experiment was performed to determine the in vivo anti-tumor effect of polydatin. Immunostaining was performed to analyze the expression of caspase-3 and Ki-67. Our results showed that polydatin inhibited cell proliferation in a concentration-dependent and time-dependent manner in the HCC cell lines. Polydatin also induced cell apoptosis in a concentration-dependent manner possibly via increasing the caspase-3 activity, and up-regulating the protein expression of caspase-3, caspase-9, Bax, and down-regulating the protein expression of Bcl-2. In addition, polydatin treatment had an inhibitory effect on cell proliferation, invasion and migration in HCC cell lines. Polydatin treatment also suppressed the Wnt/beta-catenin signaling activities in HCC cells. Polydatin treatment significantly reduced tumor growth in nude mice inoculated with HepG2 cells, suppressed the expression of Ki-67, and increased caspase-3 expression and TUNEL activity. Our data indicated the important role of polydatin for the suppression of HCC progression.


Noninvasive evaluation of bile leakage.
Three patients had internal biliary leakage demonstrated by hepatobiliary scintigraphy. In two, it was secondary to trauma and in one patient, it was related to spontaneous bile leakage of unknown causation. Although ultrasound of the abdomen is a highly accurate modality for the detection and anatomic localization of cystic collections, the ultrasonic features of a biloma are identical to those of other cystic lesions in the abdomen. In many patients, hepatobiliary and gallium scintigraphy ca help in the differentiation between biloma and abscesses. These patients stressed the need for hepatobiliary imaging, multiple projections and delayed views for the detection and localization of abnormal collections of bile. The biliary excretion of gallium should be taken into consideration when interpreting these scans.


The local anesthetic activity of tetrodotoxin alone and in combination with vasoconstrictors and local anesthetics.
Tetrodotoxin (TTX), alone and in combination with various vasoconstrictors and local anesthetics, was evaluated for its ability to produce peripheral nerve blocks in the rat and central neural blocks in the cat and dog. High frequency and long duration of block can be attained if sufficiently high concentrations of TTX are used, although latency is long and high dosage may produce systemic toxicity. Frequency and mean duration of block can be increased and systemic toxicity reduced if TTX is administered with a vasoconstrictive agent. Conventional local anesthetics also enhance the nerve-blocking activity of TTX. When appropriate concentrations of TTX and local anesthetics are used, a high frequency of blocks characterized by short latency and long duration can be demonstrated. The studies present some indirect evidence that local anesthetics enhance TTX activity by reversibly increasing the permeability of various neural barriers to TTX.


Pronounced production of polyclonal immunoglobulin G1 in the synovial fluid of goats with caprine arthritis-encephalitis virus infection.
Infection of goats with caprine arthritis-encephalitis virus, a lentivirus, resulted in arthritis characterized by the production of intrasynovial immunoglobulin G1 concentrations that were 2 to 5.3 times the serum concentrations in the inoculated carpi at 6 months postinoculation. The intrasynovial immunoglobulin was polyclonal, and its presence was accompanied by increased albumin leakage into the joints. Synovial fluid immunoglobulin levels fluctuated temporally but remained elevated compared with medium-inoculated controls for 38 months after infection. Elevated immunoglobulin G1 concentrations correlated with focal sublumenal plasmacytic infiltrates in the synovia of inoculated carpi at 5 months postinoculation. Inflammation in the uninoculated joints of infected goats was also accompanied by increased intrasynovial immunoglobulin G1 levels. Antibody to systemically administered antigens was a greater proportion of the immunoglobulin population in sera than in synovial fluids of infected goats, suggesting that antibody production to local antigens was responsible for increased intrasynovial immunoglobulin G1 levels.


Endothelial function and inflammation in coronary artery disease.
Evidence supports the central role of endothelium and inflammation in all phases of the atherosclerotic process. Clinical studies have shown their prognostic potential for the development of ischaemic events and for adverse outcome after acute coronary syndromes. Reduction in inflammatory levels and improving endothelial function by traditional and novel treatment strategies were associated with a proportional reduction in cardiovascular events. However, randomised controlled trials are required to explore further whether drugs targeting the inflammatory process and endothelial function will constitute a reasonable adjunctive treatment for patients with coronary artery disease.


End-stage renal disease and low level exposure to lead, cadmium and mercury; a population-based, prospective nested case-referent study in Sweden.
Cadmium (Cd), lead (Pb), and mercury (Hg) cause toxicological renal effects, but the clinical relevance at low-level exposures in general populations is unclear. The objective of this study is to assess the risk of developing end-stage renal disease in relation to Cd, Pb, and Hg exposure. A total of 118 cases who later in life developed end-stage renal disease, and 378 matched (sex, age, area, and time of blood sampling) referents were identified among participants in two population-based prospective cohorts (130,000 individuals). Cd, Pb, and Hg concentrations were determined in prospectively collected samples. Erythrocyte lead was associated with an increased risk of developing end-stage renal disease (mean in cases 76 μg/L; odds ratio (OR) 1.54 for an interquartile range increase, 95% confidence interval (CI) 1.18-2.00), while erythrocyte mercury was negatively associated (2.4 μg/L; OR 0.75 for an interquartile range increase, CI 0.56-0.99). For erythrocyte cadmium, the OR of developing end-stage renal disease was 1.15 for an interquartile range increase (CI 0.99-1.34; mean Ery-Cd among cases: 1.3 μg/L). The associations for erythrocyte lead and erythrocyte mercury, but not for erythrocyte cadmium, remained after adjusting for the other two metals, smoking, BMI, diabetes, and hypertension. Gender-specific analyses showed that men carried almost all of the erythrocyte lead and erythrocyte cadmium associated risks. Erythrocyte lead is associated with end-stage renal disease but further studies are needed to evaluate causality. Gender-specific analyses suggest potential differences in susceptibility or in exposure biomarker reliability.


High-sensitivity detection of carbohydrate antigen 15-3 using a gold/zinc oxide thin film surface plasmon resonance-based biosensor.
We report that gold/zinc oxide (Au/ZnO) nanocomposite films were effectively employed to enhance the performance of surface plasmon resonance (SPR) for the detection of tumor markers. Carbohydrate antigen 15.3 (CA15-3), a tumor marker for breast cancer, was chosen as a model analyte. We analyzed intensity response to the samples at various concentrations (0.0125 U/mL to 160 U/mL) in pleural fluid to evaluate the detection capability of the SPR biosensor based on Au/ZnO thin films. The linear range extended from 1 to 40 U/mL with a correlation coefficient of R(2) = 0.991 and a limit of detection reaching 0.025 U/mL at a signal-to-noise ratio of 3:1. Compared with the degree of the shift in SPR intensity induced by the specific binding event between antibody and antigen, the change of intensity on the Au/ZnO layers was increased by at least 2 fold over that on the gold/chromium (Au/Cr) layers. In addition, we determined that the Au/ZnO layers allowed for a detection limit 4 times lower than the Au/Cr layers, which are in widespread use as the sensing interfaces in current SPR-based detectors. In conclusion, the use of Au/ZnO films greatly enhanced the SPR signal yield for this bimolecular interaction and showed high sensitivity.


Ultrasensitive Detection of Ebola Virus Oligonucleotide Based on Upconversion Nanoprobe/Nanoporous Membrane System.
Ebola outbreaks are currently of great concern, and therefore, development of effective diagnosis methods is urgently needed. The key for lethal virus detection is high sensitivity, since early-stage detection of virus may increase the probability of survival. Here, we propose a luminescence scheme of assay consisting of BaGdF5:Yb/Er upconversion nanoparticles (UCNPs) conjugated with oligonucleotide probe and gold nanoparticles (AuNPs) linked with target Ebola virus oligonucleotide. As a proof of concept, a homogeneous assay was fabricated and tested, yielding a detection limit at picomolar level. The luminescence resonance energy transfer is ascribed to the spectral overlapping of upconversion luminescence and the absorption characteristics of AuNPs. Moreover, we anchored the UCNPs and AuNPs on a nanoporous alumina (NAAO) membrane to form a heterogeneous assay. Importantly, the detection limit was greatly improved, exhibiting a remarkable value at the femtomolar level. The enhancement is attributed to the increased light-matter interaction throughout the nanopore walls of the NAAO membrane. The specificity test suggested that the nanoprobes were specific to Ebola virus oligonucleotides. The strategy combining UCNPs, AuNPs, and NAAO membrane provides new insight into low-cost, rapid, and ultrasensitive detection of different diseases. Furthermore, we explored the feasibility of clinical application by using inactivated Ebola virus samples. The detection results showed great potential of our heterogeneous design for practical application.


Transient granulocyte maturation arrest: discovery by flow cytometry of a variant form of agranulocytosis.
A 5-year-old child presented with pneumonia and agranulocytosis. A Wright-stained peripheral blood smear showed only cells which had the morphological appearance of lymphocytes, plus a few monocytes and eosinophils. A bone marrow aspirate smear showed a complete lack of recognizable granulocyte precursors. However, the admission CBC and differential performed by automated flow cytochemistry (Technicon Instruments Corporation H-6000) measured 32% granulocytes as determined by peroxidase activity. Cytochemical stains on the blood and marrow smears revealed that many of the cells that had the morphological appearance of lymphocytes were positive for myeloperoxidase activity. Special studies on these cells revealed them to be abnormal, intermediate granulocytes with azurophilic, peroxidase-containing primary granules, but with few secondary granules and limited lactoferrin activity. Over 28 days the child recovered, first with granulocytic hyperplasia in the marrow and then a return of the peripheral blood to normal. This is the first case report of an episode of transient "agranulocytosis" which in reality was a maturation arrest.


Improved consistency in DNPH-mediated pyruvic acid analysis of onion juice by modifying the sample processing order.
Onion pungency is commonly measured on absorbency of the wine pink color that results from adding NaOH to the heated mixture of dinitrophenylhydrazine (DNPH) and onion juice. However, significant variation exists among several modifications of the original Schwimmer and Weston (SW) method. We observed differences in pyruvic acid concentrations of 20%-30% between our automated method and a batch method with manual absorbency readings. To determine the source of the differences, we examined the heating time and waiting time of the sample-DNPH mixtures and found no differences. The differences were caused by differential color degradation between the pyruvic acid standards and onion juice samples. These differences could be minimized by reading the absorbency within 1 min of NaOH addition. Using this information, we devised the one-by-one method to control the reading time at 30 s. We compared 5 different analysis methods of 40 onion samples representing 4 onion colors. The automated, high-performance liquid chromatography, and SW methods had similar results, with only about a 5% difference. However, the batch method resulted in differences of approximately 24% as compared to the automated method. The one-by-one method produced very comparable results, within 5%, to the automated method. By modifying the procedure to ensure a uniform and fast reading time, we increased the consistency between the pungency analysis methods. Therefore, fast and uniform absorbency reading time is essential for an accurate measurement of pungency in undiluted onion juice.


The Role of the Wnt Pathway in VEGF/Anti-VEGF-Dependent Control of the Endothelial Cell Barrier.
Investigate the contribution of the Wnt pathway to vascular endothelial growth factor (VEGF)/anti-VEGF-mediated control of endothelial cell permeability. High glucose-treated primary human retinal endothelial cells (HRECs) were exposed to either VEGF, or VEGF and then anti-VEGF. Changes in gene expression were assayed by RNAseq and qRT-PCR. Permeability was monitored by electrical cell-substrate impedance sensing (ECIS). Approaches to activate the Wnt pathway included treatment with LiCl and overexpression of constitutively activated β-catenin. β-catenin-dependent transcriptional activity was monitored in HRECs stably expressing a TCF/LEF-driven reporter. VEGF/anti-VEGF altered expression of genes encoding many members of the Wnt pathway. A subset of these genes was regulated in a way that is likely to contribute to control of the endothelial cell barrier. Namely, the VEGF-induced alteration of expression of such genes was reversed by anti-VEGF, and such adjustments occurred at times corresponding to changes in barrier function. While pharmacological and molecular approaches to activate the Wnt pathway had no effect on basal permeability, they suppressed VEGF-induced relaxation. Furthermore, anti-VEGF-mediated restoration of barrier function was unaffected by activation of the Wnt pathway. VEGF/anti-VEGF engages multiple members of the Wnt pathway, and activating this pathway enforces the endothelial barrier by attenuating VEGF-induced relaxation. These data suggest that FDA-approved agents such as LiCl may be an adjuvant to anti-VEGF therapy for patients afflicted with blinding conditions including diabetic retinopathy.


Optimum Diffraction-Corrected Frequency-Shift Estimator of the Ultrasonic Attenuation Coefficient.
The ultrasonic attenuation coefficient is an important parameter that has been studied extensively in Quantitative Ultrasound and Tissue Characterization. There are various methods described in the literature that estimate this parameter by measuring either spectral difference (i.e., decay) or spectral shift of the backscattered echo signal. Under ideal conditions, i.e., in the absence of abrupt changes in tissue backscattering, Spectral Difference methods can produce estimates with high accuracy and precision. On the other hand, diffraction-corrected Spectral Shift methods (e.g., the Hybrid method) are better suited for application in practical settings using clinical ultrasound scanners. However, current Spectral Shift methods use inefficient frequency shift estimators that ultimately degrade the quality of attenuation coefficient estimates. In this paper, a probabilistic model of the backscattered radiofrequency (RF) echo is used to derive the Cramér-Rao lower bound (CRLB) on estimation variance of the spectral centroid. Next, an efficient correlation-based shift estimator is presented that achieves the CRLB. Used in conjunction with a well-characterized reference phantom to correct for diffraction and other system-related effects, this estimator greatly improves the accuracy and precision of Spectral- Shift attenuation estimation. A theoretical analysis of this method is provided, and its performance is quantitatively compared with that of the Hybrid method using simulated and experimental phantom studies. A minimum of 3-fold reduction in the standard deviation of attenuation coefficient estimates is observed using the new method.


Prevalence of pre-diabetes in adults aged 25 - 45 years in a Durban-based clinical setting, South Africa: A retrospective study.
Due to pre-diabetes being underexplored, its prevalence was investigated in study participants aged 25-45 years in a Durban-based tertiary-level clinical setting in South Africa. The study was done using a retrospective study design. Fasting blood samples from consented patients with no previous diagnosis of diabetes and within the specified age range were collected from King Edward Hospital in Durban. The pre-diabetes diagnosis was confirmed in participants with fasting glucose concentrations between 5.6 and 6.9 mmol/L and glycated haemoglobin (HbA1c) levels between 5.7 % and 6.4 % using the American Diabetes Association (ADA) and World Health Organisation (WHO) diagnosis criteria. The study participants' characterisation was stratified according to the diagnosis criterion, age, gender and ethnicity. An alarming 68 % average pre-diabetes prevalence across ADA and WHO criteria in the Durban, eThekwini district sample population. The highest prevalence was recorded using the IFG criterion (83%) and the lowest when using the HbA1c criterion (54 %). Between the White, Black and Indian ethnic groups, the Indian group were more predisposed to pre-diabetes onset, with a prevalence of 62.7 %. If pre-diabetes management is unattended, an unprecedented increase in metabolic disorders such as Type 2 Diabetes Mellitus (T2DM) and all-cause mortality incidence can be expected. Therefore, the study reveals a window of opportunity to intensify preventative measures and mitigate the incidence of T2DM.


A fibril-like assembly of oligomers of a peptide derived from β-amyloid.
A macrocyclic β-sheet peptide containing two nonapeptide segments based on Aβ(15-23) (QKLVFFAED) forms fibril-like assemblies of oligomers in the solid state. The X-ray crystallographic structure of macrocyclic β-sheet peptide 3 was determined at 1.75 Å resolution. The macrocycle forms hydrogen-bonded dimers, which further assemble along the fibril axis in a fashion resembling a herringbone pattern. The extended β-sheet comprising the dimers is laminated against a second layer of dimers through hydrophobic interactions to form a fibril-like assembly that runs the length of the crystal lattice. The second layer is offset by one monomer subunit, so that the fibril-like assembly is composed of partially overlapping dimers, rather than discrete tetramers. In aqueous solution, macrocyclic β-sheet 3 and homologues 4 and 5 form discrete tetramers, rather than extended fibril-like assemblies. The fibril-like assemblies of oligomers formed in the solid state by macrocyclic β-sheet 3 represent a new mode of supramolecular assembly not previously observed for the amyloidogenic central region of Aβ. The structures observed at atomic resolution for this peptide model system may offer insights into the structures of oligomers and oligomer assemblies formed by full-length Aβ and may provide a window into the propagation and replication of amyloid oligomers.


Protective conditioning against GVHD and graft rejection after combined organ and hematopoietic cell transplantation.
We have performed combined organ and hematopoietic cell transplantation using a similar conditioning regimen in mice and humans. In the mouse model of MHC-mismatched combined heart and marrow transplantation, we compared conditioning of BALB/c hosts with total lymphoid irradiation (TLI: 10 doses of 240 cGy each) targeted to the spleen, lymph nodes and thymus to conditioning with a single dose of sublethal total body irradiation (TBI; 450 cGy). Conditioning also included three injections of anti-thymocyte serum (ATS), in both groups. C57BL/6 heart grafts, marrow cells and blood mononuclear cells were transplanted 24 h after the completion of irradiation. Blood mononuclear cells were added to the marrow cells to engender severe graft versus host disease (GVHD) that is present after combined organ and hematopoietic cell transplantation in humans given non-myeloablative conditioning. Both TLI and TBI conditioned groups accepted the organ grafts and became stable chimeras. However, the TBI group all died of GVHD during the 100-day observation period. The TLI group survived during the same period without clinical signs of GVHD. These hosts were tolerized to the donor organ grafts, since third party grafts were rejected rapidly when transplanted after 100 days. When NK T-cell-deficient CD1d(-/-) BALB/c hosts were used instead of wild-type hosts in the TLI/ATS conditioned group, then all hosts survived but all rejected the organ grafts and almost all failed to develop stable chimerism. None developed GVHD. Since host NK T cells were required for graft acceptance and NK T cells are activated after recognition of CD1d on antigen presenting cells, we compared heart and marrow graft survival from wild-type versus CD1d(-/-) donors after transplantation to TLI and ATS conditioned wild-type hosts. Whereas marrow and heart grafts from wild-type donors were accepted, almost all grafts from CD1d donors were rejected. Grafts from control Jalpha18(-/-) donors that were NK T cell deficient but expressed CD1d were all accepted. The results indicate that host NK T cells facilitate graft acceptance by recognizing CD1d on donor cells. We applied the TLI conditioning regimen using 10 doses of 80 cGy each and 5 doses of rabbit ATG to human recipients of HLA-matched G-CSF "mobilized" blood mononuclear cell transplants for the treatment of leukemia and lymphoma [R. Lowsky, T. Takahashi, Y.P. Liu, et al., Protective conditioning for acute graft-versus-host disease. N. Engl. J. Med. 353 (2005) 1321-1331.]. Currently more than 100 transplants have been performed, and the incidence of acute GVHD has been about 4% when both MRD and MUD transplants are combined. Almost all recipients became complete chimeras after receiving grafts that contained 2-3x10(8) CD3(+) T cells/kg. In further studies, we applied the same TLI and ATG conditioning regimen to combined kidney and G-CSF "mobilized" blood stem cell transplantation from HLA-matched sibling donors. The hematopoietic grafts in the latter protocol were selected CD34(+) cells with 1x10(6) CD3(+) T cells/kg added back to the hematopoietic cells. Preliminary results indicate that stable mixed chimerism can be achieved using this protocol allowing for complete immunosuppressive drug withdrawal without GVHD or subsequent rejection episodes. Thus, conditioning with TLI based regimens can simultaneously protect against organ graft rejection and GVHD. Levels of chimerism are dependent upon the content of donor T cells in the hematopoietic graft.


Cranial computed tomography in the diagnosis of multiple sclerosis.
A group of 202 patients with suspected, probable or definite multiple sclerosis was studied, using cranial computed tomography (CT). Atrophy alone, or in combination with white-matter and periventricular lucencies, and areas of contrast enhancement, were the main abnormal findings in 52% of patients. Atrophy was detected in 44% of patients, and its frequency and severity correlated with disease duration up to 10 years, age, and disease category. Atrophic changes in the brainstem and cerebellum could be correlated with clinical data more often than supratentorial atrophy could be correlated with features such as dementia or mood changes. Lucencies in the white matter, thought to represent areas of demyelination, were noted in 21% of patients, and only a proportion of these lesions could be correlated with clinical data, the others being clinically silent. Contrast enhancement was seen in a small proportion of white-matter lesions, and was independent of disease activity and steroid medication. Electrophysiological tests and cerebrospinal fluid analysis showed a higher yield of abnormality than CT scanning in cases with suspected or possible multiple sclerosis, though in such patients CT scanning excluded alternative cerebral atrophy. Modifications of the technique of CT scanning may improve the detection rate of white-matter lesions, thereby enhancing the value of CT as a diagnostic tool in the study of patients with multiple sclerosis.


[A hundred years of child psychiatry and changes in the governance of the youth mental health services in the Netherlands- back to the future].
Dutch child- and adolescent psychiatry (cap) is going through a complex phase now that the Youth Law has decreed that cap is to be placed under control of the municipality (i.e. local government) instead of the national government. To explain and discuss this development from a historical perspective. Information was obtained from historical sources. The municipality, i.e. local government, has played an important role in parts of youth mental health since the beginning of the 20th century. In the 1980s, however, governance was transferred to the national government, only to be passed back to the municipality in the early years of the 21st century. The level of governance, be it local or national, can bring both advantages and disadvantages to the youth mental health services. Up till now there have been several transfers from one level of governance to the other. It would be better if the two levels of governance were combined and the various parts controlled by a combination of the most appropriate types of governance.


Population Pharmacokinetics and Pharmacodynamics of the Calcimimetic Etelcalcetide in Chronic Kidney Disease and Secondary Hyperparathyroidism Receiving Hemodialysis.
Etelcalcetide is a novel calcimimetic in development for the treatment of secondary hyperparathyroidism (SHPT). A population pharmacokinetic/pharmacodynamic (PK/PD) model was developed relating etelcalcetide exposures to markers of efficacy (parathyroid hormone [PTH]) and safety (calcium) using data from three clinical studies. The semimechanistic model was developed that included allosteric activation pharmacology and understanding of calcium homeostasis. The temporal profiles for all biomarkers were well described by the model. The cooperativity constant was 4.94, confirming allosteric activation mechanism. Subjects with more severe disease (higher PTH baseline) were predicted to experience less pronounced reduction in PTH (percentage change from baseline), but more reduction in calcium (Ca; percentage change from baseline). There was no evidence that dose adjustment by any covariate was needed. Model-based simulations provided quantitative support to several elements of dosing, such as starting dose, monitoring, and titration timing for registration trials.


Primary cervical neuroblastoma in infants.
Although neuroblastoma is a relatively common malignancy of childhood, it is rarely of cervical origin. We encountered two cases of primary cervical neuroblastoma in infants. The first case was in a newborn who presented with noisy breathing, dyspnea, and choking on feeding soon after birth. Physical examination disclosed a left retropharyngeal mass near the midline causing aerodigestive compromise. Magnetic resonance imaging showed a well-defined hyperintense mass in the corresponding region. The infant underwent complete surgical excision of the mass 1 month after birth. The pathology report confirmed neuroblastoma. Horner's syndrome developed soon after surgery but resolved spontaneously within 6 months. No recurrence was noted during a 3-year follow-up period. The second patient, a 5-month-old female, presented with stridor and shortness of breath of 2-weeks' duration due to a right cervical mass. Magnetic resonance imaging showed a parapharyngeal mass suggestive of neuroblastoma. Complete surgical excision was done at the expense of sacrificing the sympathetic nerve trunk. The pathologic diagnosis was moderately differentiated neuroblastoma. No recurrence was noted when the patient was seen 2 years after surgery, although right-sided Horner's syndrome persisted. Surgical intervention can result in a good prognosis for low stage cervical neuroblastomas; however, long-term follow-up is important.


Alveolar soft-part sarcoma of the head and neck: clinical and imaging features in five cases.
Alveolar soft-part sarcoma (ASPS) of the head and neck is an extremely rare malignancy. Although the clinical and imaging features of this tumor have been reported, a periodic review of unusual tumors is useful. The purpose of this study was to describe the clinical and imaging features of ASPS of the head and neck. Between January 1990 and May 2004 at our institution, five head and neck ASPS were diagnosed in five patients (two male and three female patients; age range, 4-22 years). Clinical and imaging findings were reviewed retrospectively. Imaging studies consisted of contrast material-enhanced CT (in four patients), MR imaging (in four patients), and digital subtraction angiography (in two patients). The locations of the tumor were tongue in two cases, larynx in one case, buccal space in one case, and paravertebral space in one case. This tumor appeared as a large lobulating-contoured mass with high signal intensity and flow voids on T2-weighted images and showed strong enhancement on contrast-enhanced CT and MR images. Preoperative angiography showed high vascularity. Wide surgical excisions were performed in four cases. Mean follow-up periods were 16 months (range, 6-30 months), and no recurrence was noted except for the laryngeal case. ASPS should be included in the differential diagnosis of head and neck tumor when a slow-growing, large mass with high signal intensity and flow voids on T2-weighted images and strong enhancement on contrast-enhanced CT or MR image is seen, particularly in young female patients.


Physiology and plasticity of putative cough fibres in the Guinea pig.
Cough is initiated by activation of afferent nerve fibers with rapidly adapting receptors (RAR) that conduct action potentials in the Adelta range. In addition, various stimuli that activate airway unmylenated C-fibres evoke cough reflexes. We have used a vagally innervated, larynx-trachea-bronchus preparation, isolated from guinea pigs, to study the pharmacology of RARs and C-fibres in vitro. In this preparation afferent fibres with the RAR phenotype are exquisitely sensitive to mechanical perturbation of their receptive fields, but are unaffected by a variety of mediators (e.g. prostaglandins, histamine, bradykinin, serotonin) and by capsaicin. By contrast, C-fibres are much less sensitive to mechanical stimulation, but can be activated by capsaicin and bradykinin. Preliminary evidence supports the hypothesis that bradykinin activate C-fibre by stimulating the capsaicin (vanilloid) receptor VR1. Acids activate both C-fibres and RARs. Acids stimulate RAR fibres by a mechanism that is rapidly inactivated. C-fibres are stimulated by both a rapidly inactivating mechanism, as well as a slowly inactivating mechanism. Drugs that block VR1 inhibit the latter mechanism. Airway inflammation substantially increases the mechanical sensitivity of RAR fibres without affecting their adaptive properties. Airway inflammation also causes a phenotypic switch in neuropeptide innervation of the airways that RAR neurons begin to synthesis neurokinins and calcitonin gene related peptide. In non-inflamed animals these peptides are expressed only in C-fibre neurons. Thus, airway inflammation may not only increase the sensitivity of cough fibres, but may also qualitatively change the role played by sensory neuropeptides in cough reflexes.


Acceleration effect of basic fibroblast growth factor on the regeneration of peripheral nerve through a 15-mm gap.
In this study, nerve guides composed of poly(D,L-lactide) (PDLLA) were fabricated and used in the repair of transected sciatic nerves (15-mm gaps) of rats. Nerve guides with a two-ply structure (inner layer dense, outer layer microporous) were prepared by controlling the solvent evaporation rate. Then basic fibroblast growth factor (bFGF) was embedded in the inner layer of the nerve guides. Thus the inner dense layer not only could prevent the ingrowth of fibroblast and avoid the outgrowing nerve cable, but it also could retain the released bFGF in the guide lumen. The outer porous layer allowed vascular ingrowth and the diffusion of essential nutrients into the guide lumen. The data show that by using this nerve guide, the transected 15-mm sciatic nerve was regenerated successfully within 4 months. The recovery of function of the regenerated nerves was significantly accelerated by bFGF, as indicated by an electrostimulation test and histologic assays. In addition, the bFGF retained its bioactivity during embedding and continuously was released from the matrix, as confirmed by the results of both the dorsal root ganglia (DRG) and the Schwann cell culture in the presence of PDLLA matrix containing bFGF. The released bFGF enhanced the ability of the nerve fibers to sprout from dorsal root ganglia, and it accelerated the proliferation of Schwann cells.


In vivo optical imaging of human lymphoma xenograft using a library-derived peptidomimetic against alpha4beta1 integrin.
Increasing literature suggests that cell adhesion molecule alpha4beta1 integrin plays a pivotal role in autoimmune diseases and cancer development. Noninvasive visualization of alpha4beta1 integrin in vivo will facilitate the understanding of its involvement in disease progression and development of targeted therapies. Due to the lack of high-affinity targeting ligands, molecular imaging of alpha4beta1 integrin is much less explored than that of alphavbeta3 and alphavbeta5 integrins. We have recently reported using the one bead-one compound combinatorial library method to identify a peptidomimetic, LLP2A, that preferentially binds to activated alpha4beta1 integrin. Here, we described the use of LLP2A-Cy5.5 conjugate as an in vivo optical imaging probe in a human lymphoma xenograft model. This univalent LLP2A-Cy5.5 conjugate retained the binding activity and specificity to alpha4beta1 integrin as shown by cell binding assays using alpha4beta1-positive Molt-4 T-leukemia cells. The subcutaneous Molt-4 tumor was clearly visualized from 1 to 24 h after tail vein injection of the conjugate. Direct imaging and confocal microscopic examination of excised tumors and organs confirmed the accumulation of LLP2A in tumors and revealed very little or no uptake in normal organs except for lymph nodes. Kidney uptake was high when the whole organ was scanned but it was negative when examined microscopically, suggesting that LLP2A bound to the renal tubules loosely. Tumor uptake of LLP2A-Cy5.5 conjugate was blocked by excess unlabeled LLP2A. This study showed that the combinatorial chemical library-derived peptidomimetic LLP2A can be easily developed into an optical imaging probe for noninvasively monitoring of activated alpha4beta1 integrin in vivo.


Freeze-fracture studies of plasma membranes of astrocytes in freezing lesions.
Epileptic foci are frequently associated with a proliferation and hypertrophy of fibrous astrocytes. Such proliferating astrocytes, which are involved in scar formation and are referred to as reactive, have been characterized by a number of morphological changes involving the cytoplasmic organelles and nucleus. With the development of freeze-fracture techniques, which allow for the splitting and analysis of the macromolecular structure of biological membranes, another morphological characteristic could be used to define reactive astrocytes. This characteristic is the alteration in the normal number and distribution of intramembranous particles. Specifically, there is an increase in the number of orthogonal arrays of intramembranous particles, called assemblies, in the plasma membrane of reactive astrocytes. Besides the presence of assemblies in the plasma membranes of astrocytes, similar particle arrays are also found in the central nervous system (CNS), in cell membranes of ependymal cells, in Müller cells near the inner limiting membrane of the retina, and in satellite cells in spinal ganglia. Assemblies are concentrated in the plasma membranes of astrocytic processes that form the glia limitans at the outer surface of the brain and the perivascular sheath around parenchymal blood vessels in the adult mammalian CNS. Plasma membranes of perineuronal astrocytic processes and astrocytic cell bodies have few, if any, assemblies. The factor responsible for this polarity of assemblies in the normal CNS is unknown. However, this polarity is lost when astrocytes respond to various types of injuries to the CNS. It has been postulated that gliosis, resulting in an epileptic scar, may be a response of cortical astrocytes to changes in the local cellular environment and that this reactivity of astrocytes may be related to plasma membrane changes. However, little is known about the initiation of the astrocytic response or the involvement of the astrocytic plasma membrane. Because of the presence of assemblies, astrocytic plasma membranes are an ideal subject for a morphological investigation of alterations of the cell membrane. Assemblies have a distinctive structure and changes in their number and distribution can be readily identified. Also, since there is polarity of the assemblies in astrocytic plasma membranes, such that their number is greatest in subpial and perivascular areas and least in the perineuronal regions, it is easy to identify when assemblies are added or lost from the astrocytic plasma membranes of a particular brain region.(ABSTRACT TRUNCATED AT 400 WORDS)


Chromosomal deletion and rearrangement in Streptomyces glaucescens.
The Streptomyces glaucescens genome frequently undergoes gross genomic rearrangement events which result in the deletion of extremely large segments of chromosomal DNA. The structure and origin of the DNA forming the novel junctions arising from five of these deletion events are described. Only one junction proved to be the result of a relatively simple event; the remainder were more complex, with one involving DNA which originated from at least five distinct loci. In three of the investigated cases, DNA sequences present in the junctions appeared to have resulted from the duplication of previously unique sequences, suggesting that duplication of chromosomal segments may be an important factor in genetic instability. The nucleotide sequences surrounding these junctions and their respective wild-type termini were determined.


Evolutionary history and ecology shape the diversity and abundance of phytochemical arsenals across monkeyflowers.
We examine the extent to which phylogenetic effects and ecology are associated with macroevolutionary patterns of phytochemical defence production across the Mimulus phylogeny. We grew plants from 21 species representing the five major sections of the Mimulus phylogeny in a common garden to assess how the arsenals (NMDS groupings) and abundances (concentrations) of a phytochemical defence, phenylpropanoid glycosides (PPGs), vary across the phylogeny. Very few PPGs are widespread across the genus, but many are common to multiple sections of the genus. Phytochemical arsenals cluster among sections in an NMDS and are not associated with total concentration of PPGs. There is a strong phylogenetic signal for phytochemical arsenal composition across the Mimulus genus, whereas ecological variables such as growing season length, latitude, and elevation do not significantly influence arsenal. In contrast, there is little phylogenetic signal for total PPG concentration, and this trait is significantly influenced by several ecological factors. Phytochemical arsenals and abundances are influenced by plant life history form. Both phylogenetic effects and ecology are related to phytochemical patterns across species, albeit in different ways. The independence of phytochemical defence concentrations from arsenal compositions indicates that these aspects of defence may continue to evolve independently of one another.


An amperometric method for the detection of amitrole, glyphosate and its aminomethyl-phosphonic acid metabolite in environmental waters using passive samplers.
The herbicides amitrole and glyphosate, and its metabolite aminomethyl-phosphonic acid (AMPA), in water samples have been directly analysed by high-performance liquid chromatography using an electrochemical (EC) detector. Limits of detection of 0.3 microg mL(-1) for glyphosate, 0.05 microg mL(-1) for AMPA and 0.03 microg mL(-1) for amitrole were comparable to those obtained by other authors using EC and also by liquid chromatography coupled to mass spectrometry, but the latter method requires derivatisation and pre-concentration of the sample whereas EC methods show similar sensitivity without the need of any derivatisation. The method was specifically designed to analyse extracts from passive samplers used for monitoring of polar herbicide residues in waters. To this purpose, three types of Empore disks were tested for their ability to adsorb and desorb these ionic, polar analytes. A procedure for their extraction from the membranes and reducing the interferences from other substances present in natural waters (i.e. humic acids) is described. The method is simple, does not require sophisticated equipment and is valid for the analysis and monitoring of herbicides residues using passive samplers.


Discontinuation and nonpublication of nasopharyngeal carcinoma clinical trials.
To determine the extent of research waste in the field of nasopharyngeal carcinoma (NPC). In this cross-sectional study, we explored the rates, causes and predictors of discontinuation and nonpublication of NPC clinical trials. The sample was derived using the ClinicalTrials.gov advanced search function. Adjusted logistic regression was used to ascertain the effect of trial characteristics on completion and publication status. If a trial discontinuation explanation or publication status could not be determined through the systematic search, the corresponding author was emailed. Ultimately, 311 NPC clinical trials were included (255 [82.0 %] completed and 56 [18.0 %] discontinued trials). The most common reason for trial discontinuation was poor accrual (50 %, 23/46). Industry funding (adjusted OR, 3.12; P = 0.003) and recurrent/metastatic setting (adjusted OR, 11.95; P = 0.003) were significantly associated with increased likelihood of trial discontinuation. Of the 207 completed trials included in the publication query, 141 (68.1 %) were published in peer-reviewed journals, 10 (4.8 %) had results only available on ClinicalTrials.gov, and 56 (27.1 %) remained unpublished 3 or more years after trial completion. Radiation with or without pharmacologic interventions significantly increased the potential of publication (adjusted OR, 3.20; P = 0.048). Among published trials, the median time to publication was 28.47 months (interquartile range, 15.27-44.98 months). We identified the difficulties inherent in NPC clinical trials from completion to publication. This represents considerable research waste in NPC, thus raising ethical concerns about the concealment of clinical data and futile patient participation and attendant risks.


[New distribution of tick-borne diseases in temperate countries].
Since 20 or 30 years, emerging tick-borne diseases are regularly reported. Recent data show that at the same time, we have to expect a change in distribution of some disease as the European tick-borne encephalitis, due to the strong relation between ticks cycle and temperatures for the transmission of this virus.


Do repeated risk factor measurements influence the impact of education on cardiovascular mortality?
It has been questioned if the excess cardiovascular disease (CVD) mortality by lower educational level can be fully explained by conventional modifiable CVD risk factors. Our objective was to examine whether repeated measures over time of risk factors (smoking, physical inactivity, blood pressure, total cholesterol and body mass index) explain more of the socioeconomic gradient in CVD mortality than if they are measured only once. A cohort of 34 884 men and women attended all three screenings (1974-1978, 1977-1983 and 1985-1988) in the Norwegian Counties Study and were followed for CVD mortality through 2009 by linkage to the Norwegian Cause of Death Registry. Age-adjusted and sex-adjusted HR of CVD mortality was 2.32 (95% CI 1.93 to 2.80) for basic relative to tertiary educated individuals. The HR was attenuated by 48% (HR 1.54 (1.28 to 1.87)) when adjusted for CVD risk factors measured at baseline and by 56% (HR 1.45 (1.20 to 1.75)) when two repeated measurements ascertained 5 years apart were added to the model. Similarly, absolute risk difference in CVD mortality by education was attenuated by 62% when adjusted for baseline and by 72% when adjusted for repeated measurements of risk factors. In this cohort, repeated measurements of risk factors seemed to explain more of the educational gradient in CVD mortality. This suggests that a substantial part of the excess CVD mortality among those with lower education might be explained by conventional risk factors.


Spermidine oxidase-derived H₂O₂ regulates pollen plasma membrane hyperpolarization-activated Ca(2+) -permeable channels and pollen tube growth.
Spermidine (Spd) has been correlated with various physiological and developmental processes in plants, including pollen tube growth. In this work, we show that Spd induces an increase in the cytosolic Ca(2+) concentration that accompanies pollen tube growth. Using the whole-cell patch clamp and outside-out single-channel patch clamp configurations, we show that exogenous Spd induces a hyperpolarization-activated Ca(2+) current: the addition of Spd cannot induce the channel open probability increase in excised outside-out patches, indicating that the effect of Spd in the induction of Ca(2+) currents is exerted via a second messenger. This messenger is hydrogen peroxide (H₂O₂), and is generated during Spd oxidation, a reaction mediated by polyamine oxidase (PAO). These reactive oxygen species trigger the opening of the hyperpolarization-activated Ca(2+) -permeable channels in pollen. To provide further evidence that PAO is in fact responsible for the effect of Spd on the Ca(2+) -permeable channels, two Arabidopsis mutants lacking expression of the peroxisomal-encoding AtPAO3 gene, were isolated and characterized. Pollen from these mutants was unable to induce the opening of the Ca(2+) -permeable channels in the presence of Spd, resulting in reduced pollen tube growth and seed number. However, a high Spd concentration triggers a Ca(2+) influx beyond the optimal, which has a deleterious effect. These findings strongly suggest that the Spd-derived H₂O₂ signals Ca(2+) influx, thereby regulating pollen tube growth.


An increase in the carbohydrate moiety of alpha 2-macroglobulin is associated with systemic lupus erythematosus (SLE).
Using lectin blots in conjunction with peptide mapping, alpha 2-macroglobulin micropurified from systemic lupus erythematosus (SLE) patients was shown to become abnormally glycosylated suggesting the occurrence of complex glycosylation in this pathological condition. To confirm there is indeed a quantitative increase in specific monosaccharides in this protein; alpha 2-macroglobulin was micropurified from a battery of 37 serum samples which included 6 normal donors (3 male and 3 female), 23 SLE patients, 6 rheumatoid arthritis patients, 1 mixed connective tissue disease patient, and 1 Sjogren's syndrome patient; for carbohydrate analysis. It was noted that the concentration of total monosaccharides in alpha 2-macroglobulin micropurified from serum samples of SLE patients is significantly higher than normal donors with a mean +/- SD of 188 +/- 410 micrograms/mg protein (SLE, n = 23) versus 14.5 +/- 4 micrograms/mg protein (normal, n = 6) even though there was a high variation in the level of monosaccharides among the SLE patients. An increase in oligosaccharides in alpha 2-macroglobulin from SLE patients compared to normal subjects was confirmed by concanavalin A (Con A) blots using peptide fragments derived from the micropurified protein. Since the interaction of peptide fragments derived from alpha 2-macroglobulin with Con A requires the presence of mannose and/or glucose residues, we have also examined if there are any correlations between the levels of mannose and glucose in alpha 2-macroglobulin and SLE. The concentration of mannose (38 +/- 60 micrograms/mg protein) in alpha 2-macroglobulin derived from SLE patients was significantly higher than normal donors (mannose, 4.8 +/- 1 micrograms/mg protein) however, the concentration of glucose in alpha 2-macroglobulin derived from SLE patients when compared to normal donors was not statistically significant, 18 +/- 20 micrograms/mg protein in SLE versus 2 +/- 0.5 micrograms/mg protein in normal donors due to high variation between samples. Also, the concentration of galactose in alpha 2-macroglobulin from SLE patients was significantly higher than normal donors (45.7 +/- 173 micrograms/mg protein versus 0.13 +/- 0.03 microgram/mg protein). These results illustrate quantification of carbohydrate in selected glycoproteins such as alpha 2-macroglobulin may be a novel and alternative clinical marker for SLE.


Preoperative left hepatic lobe devascularisation to minimize perioperative bleeding in a Jehovah's Witness undergoing left hepatectomy.
Major liver resection in a Jehovah's Witness presents unique clinical challenges requiring multimodal blood minimization strategies to reduce perioperative complications. We report a case where complete left hepatic lobe devascularisation was undertaken to minimize bleeding in a Jehovah's Witness undergoing left hepatectomy. A 65-year-old male Jehovah's Witness presented for open left hepatectomy for a large left-sided hepatocellular carcinoma involving segment IV of the liver. Three weeks prior to surgery, the patient underwent left portal vein embolization. To isolate and devascularise the left lobe, the gastroduodenal artery and left hepatic artery were then occluded with coils. The bed of the left hepatic artery was then embolised to stasis with particles. Finally, the anastomosis back to the right hepatic artery was also occluded by coils. The patient underwent uneventful surgery with an estimated blood loss of 450mls. Left hepatectomy in a Jehovah's Witness patient is feasible but requires careful planning and a multidisciplinary approach. Major liver resection represents a well defined but complex haemostatic challenge from tissue and vascular injury, further complicated by hepatic dysfunction, and activation of inflammatory, haemostatic and fibrinolytic pathways. In addition to the haemoglobin optimization strategies utilized preoperatively, the use of interventional radiology techniques to further reduce perioperative bleeding should be considered in all complex cases. Combination of portal vein embolization and hepatic lobe devascularisation to produce total vascular occlusion of inflow to the left lobe radiologically allowed a near bloodless surgical field during major liver resection in a Jehovah's Witness patient.


Facial expression training optimises viewing strategy in children and adults.
This study investigated whether training-related improvements in facial expression categorization are facilitated by spontaneous changes in gaze behaviour in adults and nine-year old children. Four sessions of a self-paced, free-viewing training task required participants to categorize happy, sad and fear expressions with varying intensities. No instructions about eye movements were given. Eye-movements were recorded in the first and fourth training session. New faces were introduced in session four to establish transfer-effects of learning. Adults focused most on the eyes in all sessions and increased expression categorization accuracy after training coincided with a strengthening of this eye-bias in gaze allocation. In children, training-related behavioural improvements coincided with an overall shift in gaze-focus towards the eyes (resulting in more adult-like gaze-distributions) and towards the mouth for happy faces in the second fixation. Gaze-distributions were not influenced by the expression intensity or by the introduction of new faces. It was proposed that training enhanced the use of a uniform, predominantly eyes-biased, gaze strategy in children in order to optimise extraction of relevant cues for discrimination between subtle facial expressions.


Systematic single cell RNA sequencing analysis reveals unique transcriptional regulatory networks of Atoh1-mediated hair cell conversion in adult mouse cochleae.
Regeneration of mammalian cochlear hair cells (HCs) by modulating molecular pathways or transcription factors is a promising approach to hearing restoration; however, immaturity of the regenerated HCs in vivo remains a major challenge. Here, we analyzed a single cell RNA sequencing (scRNA-seq) dataset during Atoh1-induced supporting cell (SC) to hair cell (HC) conversion in adult mouse cochleae (Yamashita et al. (2018)) using multiple high-throughput sequencing analytical tools (WGCNA, SCENIC, ARACNE, and VIPER). Instead of focusing on differentially expressed genes, we established independent expression modules and confirmed the existence of multiple conversion stages. Gene regulatory network (GRN) analysis uncovered previously unidentified key regulators, including Nhlh1, Lhx3, Barhl1 and Nfia, that guide converted HC differentiation. Comparison of the late-stage converted HCs with the scRNA-seq data from neonatal mouse cochleae (Kolla et al. (2020)) revealed that they closely resemble postnatal day 1 wild-type OHCs, in contrast to other developmental stages. Using ARACNE and VIPER, we discovered multiple key regulators likely to promote conversion to a more mature OHC-like state, including Zbtb20, Nfia, Zmiz1, Gm14418, Bhlhe40, Six2, Fosb and Klf9. Our findings provide insights into the regulation of HC regeneration in adult mammalian cochleae in vivo and demonstrate an approach for analyzing GRNs in large scRNA-seq datasets.


Data Quality Issues With Physician-Rating Websites: Systematic Review.
In recent years, online physician-rating websites have become prominent and exert considerable influence on patients' decisions. However, the quality of these decisions depends on the quality of data that these systems collect. Thus, there is a need to examine the various data quality issues with physician-rating websites. This study's objective was to identify and categorize the data quality issues afflicting physician-rating websites by reviewing the literature on online patient-reported physician ratings and reviews. We performed a systematic literature search in ACM Digital Library, EBSCO, Springer, PubMed, and Google Scholar. The search was limited to quantitative, qualitative, and mixed-method papers published in the English language from 2001 to 2020. A total of 423 articles were screened. From these, 49 papers describing 18 unique data quality issues afflicting physician-rating websites were included. Using a data quality framework, we classified these issues into the following four categories: intrinsic, contextual, representational, and accessible. Among the papers, 53% (26/49) reported intrinsic data quality errors, 61% (30/49) highlighted contextual data quality issues, 8% (4/49) discussed representational data quality issues, and 27% (13/49) emphasized accessibility data quality. More than half the papers discussed multiple categories of data quality issues. The results from this review demonstrate the presence of a range of data quality issues. While intrinsic and contextual factors have been well-researched, accessibility and representational issues warrant more attention from researchers, as well as practitioners. In particular, representational factors, such as the impact of inline advertisements and the positioning of positive reviews on the first few pages, are usually deliberate and result from the business model of physician-rating websites. The impact of these factors on data quality has not been addressed adequately and requires further investigation.


[Psychological wellbeing in the hospital administrative personnel].
Job satisfaction and psychological well being of health care workers significantly influence the quality of care. The aim of this study was to evaluate job satisfaction and psychological well-being in the administrative personnel of a large hospital in Rome-Italy. The personnel self-completed two anonymous questionnaires: the General Health Questionnaire (GHQ-12) for minor psychiatric non psychotic diseases and a validated job satisfaction questionnaire. Descriptive analysis and multiple logistic regression analysis were performed. Response rate was 65%. 74 were the questionnaires analyzed. 37.8% of respondents showed a high likelihood to develop symptoms of depression/anxiety (OR = 3.57; 95% CI 1.17-10.85). Major determinants of job dissatisfaction were the lower autonomy and poor communication in the working Unit. Factors associated with overall dissatisfaction were: insufficient training opportunities (p = 0.033), low autonomy (p = 0.023), poor communication between hospital Units (p = 0.009), unclear aims of their working Unit (p = 0.029), low agreement between personal expectations and required competence (p < 0.001). Among administrative staff job dissatisfaction was associated with high likelihood of depression/anxiety, low autonomy and poor communication, independently of sex, age, years of employment. Possible targets to improve satisfaction of administrative personnel were identified.


Nuclear export inhibitors avert progression in preclinical models of inflammatory demyelination.
Axonal damage has been associated with aberrant protein trafficking. We examined a newly characterized class of compounds that target nucleo-cytoplasmic shuttling by binding to the catalytic groove of the nuclear export protein XPO1 (also known as CRM1, chromosome region maintenance protein 1). Oral administration of reversible CRM1 inhibitors in preclinical murine models of demyelination significantly attenuated disease progression, even when started after the onset of paralysis. Clinical efficacy was associated with decreased proliferation of immune cells, characterized by nuclear accumulation of cell cycle inhibitors, and preservation of cytoskeletal integrity even in demyelinated axons. Neuroprotection was not limited to models of demyelination, but was also observed in another mouse model of axonal damage (that is, kainic acid injection) and detected in cultured neurons after knockdown of Xpo1, the gene encoding CRM1. A proteomic screen for target molecules revealed that CRM1 inhibitors in neurons prevented nuclear export of molecules associated with axonal damage while retaining transcription factors modulating neuroprotection.


The evolution of trade-offs under directional and correlational selection.
Using quantitative genetic theory, we develop predictions for the evolution of trade-offs in response to directional and correlational selection. We predict that directional selection favoring an increase in one trait in a trade-off will result in change in the intercept but not the slope of the trade-off function, with the mean value of the selected trait increasing and that of the correlated trait decreasing. Natural selection will generally favor an increase in some combination of trait values, which can be represented as directional selection on an index value. Such selection induces both directional and correlational selection on the component traits. Theory predicts that selection on an index value will also change the intercept but not the slope of the trade-off function but because of correlational selection, the direction of change in component traits may be in the same or opposite directions. We test these predictions using artificial selection on the well-established trade-off between fecundity and flight capability in the cricket, Gryllus firmus and compare the empirical results with a priori predictions made using genetic parameters from a separate half-sibling experiment. Our results support the predictions and illustrate the complexity of trade-off evolution when component traits are subject to both directional and correlational selection.


Activation of adenovirus promoters by the adenovirus E1A protein in cell-free extracts.
The primary product of the adenovirus E1A gene is a protein that is sufficient for controlling host-cell proliferation and immortalizing primary rodent cells. The mechanism by which the protein induces these cellular effects is poorly understood, but might be linked to its ability to regulate RNA transcription from a number of viral and cellular genes. The mechanism of E1A's transcriptional-activation (trans-activation) was studied here by monitoring the protein's effect on specific adenovirus promoters in two types of transcriptional systems in vitro. One of these systems consisted of extracts from transformed cells constitutively expressing E1A, and the other consisted of extracts of HeLa cells supplemented with a plasmid-encoded E1A protein purified from Escherichia coli. The results show that the E1A protein specifically stimulates transcription from adenovirus promoters; thus, the induction of cellular transcription factors is not necessary to explain the stimulation of transcription by E1A.


The effect of lamina propria on the growth and differentiation of urothelial cells in vitro.
The effect of lamina propria on the growth and differentiation of the mouse urinary bladder urothelial cells in vitro has been studied by light and electron microscopy using morphological and immunohistochemical methods. Three different types of urothelial cultures were maintained on a porous membrane in serum-free medium for 10 days. Our results showed that urothelial cells in culture types I and II were organised as the multilayer epithelium showing terminal differentiation, from the porous membrane toward the surface. On the surface of culture type I and II, superficial urothelial cells were highly differentiated. They formed tight junctions and had plaques of an asymmetric unit membrane, a hallmark of terminal differentiation in bladder superficial cells. These cultured superficial cells were also cytokeratin 20 positive. On the contrary, urothelial cells in culture type III have shown a limited capacity for survival and have grown as poorly differentiated monolayer. These results revealed that reciprocal intercellular signalling between the lamina propria and urothelial cells is essential to achieve terminal differentiation of urothelial cells in vitro.


The Role of Stents as Bridge to Surgery for Acute Left-Sided Obstructive Colorectal Cancer: Meta-Analysis of Randomized Controlled Trials.
The role of self-expanding metallic stents (SEMS) as a bridge to surgery for acute left-sided obstructive colorectal cancer has remained controversial. To study the efficacy of this approach, we performed a meta-analysis at the gastrointestinal surgery center of Sichuan Academy of Medical Sciences and Sichuan Provincial People's Hospital (Eastern Hospital). Two PubMed and science-direct electronic databases were searched up to December 30, 2017. Eligible studies were randomized controlled trials (RCTs). We selected 8 RCTs articles, which included 497 cases. The directly stoma rates were significantly lower in the stent group (odds ratio [OR] = 0.46, 95% confidence intervals [CIs] = 0.30-0.70, p = 0.0003). The successful primary anastomosis rates were significantly higher in the stent group (OR = 2.29, 95% CIs = 1.52-3.45, p < 0.0001). The post-procedural complication rates were significantly lower in the stent group (OR = 0.39, 95% CI = 0.18-0.82, p = 0.01). However, tumor recurrence rates were significantly higher in the stent group (OR = 1.79, 95% CI = 1.09-2.93, p = 0.02). Conclusions This meta-analysis confirms that SEMS placement could reduce direct stoma rate and increases the successful primary anastomosis rate; however, it was associated with a seemingly higher incidence of tumor recurrence.


No seasonal variation in physical activity of Han Chinese living in Beijing.
Physical activity (PA) is widely acknowledged to be beneficial to health and wellbeing, and is potentially influenced by a variety of environmental factors such as ambient temperature, weather conditions and air pollution levels. Since these factors vary seasonally, physical activity participation may also respond seasonally. Current population studies to profile physical activity often sample individuals only once, and this may result in biased estimates if there is strong seasonal variation. We conducted a study of 40 Han Chinese adults living in Beijing using GT3X accelerometers. We measured PA levels every two months across a complete year, while simultaneously monitoring ambient temperatures and air pollution levels. Average hourly vector magnitude (VM) and percentage time spent at each PA intensity (sedentary to light, moderate, vigorous and very vigorous) were measured. General Linear models (GLMs) were used to explore the effects of time of day, temperature and PM 2.5 levels on PA. One way ANOVA was used to test whether there were seasonal differences in body weight and body fatness. The main factors influencing activity levels were the time of day and individual characteristics including age and body fatness, but there was no significant difference between the months. In addition, there was no significant impact of either ambient temperature or air pollution levels (PM2.5). There were also no significant differences over the year in the time spent at sedentary-light, moderate and very vigorous PA levels, but for vigorous PA level which occupied less than 0.5% daily physical activity, both month and individual were significant factors. The relatively constant pattern of urban daily life, independent of time of year, may override the potential impacts of environmental factors that would be anticipated to impact PA levels. These subjects did not specifically avoid activity coincident with elevated air pollution levels (PM2.5). Single week long measurements of physical activity could provide a representative measurement of the physical active levels in this population.


Elevated levels of MIC-1/GDF15 in the cerebrospinal fluid of patients are associated with glioblastoma and worse outcome.
For patients with brain tumors identification of diagnostic and prognostic markers in easy accessible biological material, such as plasma or cerebrospinal fluid (CSF), would greatly facilitate patient management. MIC-1/GDF15 (growth differentiation factor 15) is a secreted protein of the TGF-beta superfamily and emerged as a candidate marker exhibiting increasing mRNA expression during malignant progression of glioma. Determination of MIC-1/GDF15 protein levels by ELISA in the CSF of a cohort of 94 patients with intracranial tumors including gliomas, meningioma and metastasis revealed significantly increased concentrations in glioblastoma patients (median, 229 pg/ml) when compared with control cohort of patients treated for non-neoplastic diseases (median below limit of detection of 156 pg/ml, p < 0.0001, Mann-Whitney test). However, plasma MIC-1/GDF15 levels were not elevated in the matching plasma samples from these patients. Most interestingly, patients with glioblastoma and increased CSF MIC-1/GDF15 had a shorter survival (p = 0.007, log-rank test). In conclusion, MIC-1/GDF15 protein measured in the CSF may have diagnostic and prognostic value in patients with intracranial tumors.


Low-grade isthmic spondylolisthesis treated with instrumented posterior lumbar interbody fusion in U.S. servicemen.
The existing literature lacks a functional outcomes study addressing instrumented posterior lumbar fusion surgery for isthmic spondylolisthesis in physically active patients. Presently, spinal surgeons can provide only anecdotal advice when discussing operative outcomes with these patients. This is a nonrandomized analysis of consecutive military servicemen treated operatively for chronic back pain and low-grade isthmic spondylolisthesis with single-level lumbar disc degeneration with emphasis on functional outcomes. The purpose was to evaluate patient-assessed function/pain/satisfaction and military job performance in U.S. servicemen treated with posterior lumbar interbody fusion (PLIF). Thirty consecutive U.S. military servicemen with chronic low back pain and low-grade lumbar isthmic spondylolisthesis were referred to the same surgeon at a military treatment facility. All servicemen were treated operatively with instrumented PLIF using autogenous iliac crest bone graft, one or two nonthreaded interbody cages (Brantigan or Harms), and a four-pedicle screw/rod construct. A concomitant bilateral posterolateral fusion was performed in all 30 cases. The average follow-up time was 15 months (range 12-48 months). Twenty-five of the 30 servicemen completed a functional outcomes questionnaire (American Academy of Orthopaedic Surgeons/Scoliosis Research Society) with emphasis on pre- and posttreatment function, pain, and satisfaction. The servicemen were also evaluated using standard military job performance parameters. Three of the 30 servicemen (10%) requested and received a disability discharge from the military for back pain that continued throughout the postoperative period and prevented return to military duty. An additional 8 of the original 30 soldiers (27%) required some form of permanent physical activity limitation (situps/pushups/running/lifting) to permit their return to military duty, and 19 of 30 (63%) soldiers were able to return to full and unrestricted military duty after surgery. Those soldiers who were able to return to unrestricted military duty (n = 19) did so at an average of 6 months post treatment (range 2-16 months). Of the soldiers who were able to return to military duty, 21 of 30 (70%) were able to complete the posttreatment military physical fitness test at an average of 8 months postoperatively (range 2-32 months). No significant differences were observed between premorbid and postsurgical physical fitness test scores. There was a trend toward lower postsurgical scores. Complications included dural tear (n = 4), unilateral transient lower extremity paresthesia (n = 1), and wound seroma requiring reoperation (n = 1). In this nonrandomized study of 30 U.S. servicemen with chronic low back pain, low-grade isthmic spondylolisthesis, and single-level lumbar disc degeneration, instrumented PLIF surgery was associated with a high rate of return to functional military duty. Outcomes with respect to posttreatment pain, function, and satisfaction were high in patients treated with instrumented PLIF.


Conservation resource allocation, small population resiliency, and the fallacy of conservation triage.
Some conservation prioritization methods are based on the assumption that conservation needs overwhelm current resources and not all species can be conserved; therefore, a conservation triage scheme (i.e., when the system is overwhelmed, species should be divided into three groups based on likelihood of survival, and efforts should be focused on those species in the group with the best survival prospects and reduced or denied to those in the group with no survival prospects and to those in the group not needing special efforts for their conservation) is necessary to guide resource allocation. We argue that this decision-making strategy is not appropriate because resources are not as limited as often assumed, and it is not evident that there are species that cannot be conserved. Small population size alone, for example, does not doom a species to extinction; plants, reptiles, birds, and mammals offer examples. Although resources dedicated to conserving all threatened species are insufficient at present, the world's economic resources are vast, and greater resources could be dedicated toward species conservation. The political framework for species conservation has improved, with initiatives such as the UN Sustainable Development Goals and other international agreements, funding mechanisms such as The Global Environment Facility, and the rise of many nongovernmental organizations with nimble, rapid-response small grants programs. For a prioritization system to allow no extinctions, zero extinctions must be an explicit goal of the system. Extinction is not inevitable, and should not be acceptable. A goal of no human-induced extinctions is imperative given the irreversibility of species loss.


Prevalence and correlates of hazardous alcohol consumption and binge drinking among men who have sex with men (MSM) in San Francisco.
To describe heavy alcohol use patterns and correlates in a diverse sample of MSM. We used respondent-driven sampling (RDS) to enroll 252 alcohol-using MSM in San Francisco from March 2015-July 2017. We examined heavy alcohol use patterns and conducted RDS-adjusted multivariable analyses to characterize correlates of hazardous alcohol consumption and binge drinking. RDS-adjusted prevalence of weekly and at least weekly binge drinking was 24.9% and 19.3%, respectively. Hazardous consumption was common; prevalence of mid- and high-levels of hazardous drinking was 11.4% and 29.9%, respectively. In multivariable analyses, identifying as Hispanic/Latino or mixed/other race; being moderately or extremely interested in reducing alcohol use; ever receiving alcohol treatment; using ecstasy; reporting syphilis diagnosis; and having more than 5 male partners were independently associated with hazardous alcohol consumption. Less hazardous consumption was associated with having a bachelor's degree or completing post-graduate studies; and not being in a relationship. Reporting chlamydia infection; being somewhat, moderately or extremely interested in reducing alcohol use; and having multiple male sex partners were associated with higher odds of at least weekly binge drinking. Lower odds of binge drinking were associated with completing post-graduate studies. Moreover, for the outcomes of hazardous alcohol consumption and binge-drinking, we observed significant interaction effects between race/ethnicity and interest in reducing alcohol, past receipt of alcohol treatment, use of ecstasy, syphilis diagnosis, and number of male partners. Among alcohol-using MSM in San Francisco, heavy drinking patterns were common and independently associated with greater number of male sexual partners and sexually transmitted infections (STI). Moreover, significant racial/ethnic and socioeconomic disparities related to heavy alcohol use were observed and race/ethnicity modified the effect of the risk factors associated with these outcomes. These findings underscore the need to develop more MSM-specific interventions that jointly address heavy alcohol use and HIV/STI risk, as well as culturally-tailored and targeted strategies to alleviate health disparities.


Assessing the efficacy of cancer screening.
Population-based cancer screening has been established for several types of cancer in Australia and internationally. Screening may perform differently in practice from randomised controlled trials, which makes evaluating programs complex. We discuss how to assess the evidence of benefits and harms of cancer screening, including the main biases that can mislead clinicians and policy makers (such as volunteer, lead-time, length-time and overdiagnosis bias). We also discuss ways in which communication of risks can inform or mislead the community. The evaluation of cancer screening programs should involve balancing the benefits and harms. When considering the overall worth of an intervention and allocation of scarce health resources, decisions should focus on the net benefits and be informed by systematic reviews. Communication of screening outcomes can be misleading. Many messages highlight the benefits while downplaying the harms, and often use relative risks and 5-year survival to persuade people to screen rather than support informed choice. An evidence based approach is essential when evaluating and communicating the benefits and harms of cancer screening, to minimise misleading biases and the reliance on intuition.


Methods for the assessment of PTCA success.
Numerous criteria for the assessment of the immediate and late results of percutaneous transluminal coronary angioplasty (PTCA) are currently in use. Here, the values and limitations of the transstenotic gradient (TG), visual qualitative angiogram assessment, quantitative coronary angiography and the coronary flow reserve (CFR) will be discussed. Although measurement of the TG may be clinically useful, current data suggest that it does not accurately reflect the "true" pressure-flow-resistance across coronary lesions. Furthermore, the widely applied method of visual interpretation of coronary angiograms is hampered by several serious shortcomings: large intra- and interobserver variabilities, and lack of correlation with pathologic as well as intraoperative findings. In contradistinction, CFR and minimal luminal cross-sectional area (MLCA) appear to be more reliable parameters for judging the physiologic importance of (residual) coronary obstructions. In fact, given the curvilinear relation between CFR and MLCA (r = 0.92), the available evidence suggests that at the moment quantitative coronary angiography--with measurement of the MLCA immediately after PTCA--is the method of choice in assessing the efficacy of coronary angioplasty.


Activation of integrin and ceramide signalling pathways can inhibit the mitogenic effect of insulin-like growth factor I (IGF-I) in human breast cancer cell lines.
Cell counting, cell cycle analysis and Western immunoblotting were used to examine the effects of non-apoptotic doses of a ceramide analogue, C2, and a synthetic arginine-glycine-aspartic acid (RGD)-containing peptide, RGD, in MCF-7 and T47D cells to determine whether activation of these signalling pathways could alter the mitogenic potential of insulin-like growth factor I (IGF-I). IGF-I alone increased total cell number in both cell lines, associated with a rise in the percentage of cells in the S-phase of the cell cycle and a co-incident increase in cyclin A production. Treatments alone had no effects on cell number or cyclin A production relative to controls. C2 inhibited IGF-I-induced mitogenesis in both lines, whereas RGD was only effective in the T47D line. Despite inhibition of cell proliferation, IGF-I stimulation of cells in S-phase and of cyclin A levels were unaffected; however, an IGF-I-induced increase in cyclin B1 levels was inhibited by 30%. Low-dose induction of integrin and ceramide signalling pathways causes cells to be blocked in S-phase, thereby inhibiting the normal cycle of events associated with the IGF-I-induced mitotic signal. Activating these pathways may not only restrict tumour growth by induction of apoptosis but they may also directly inhibit IGF-I-induced cell proliferation.


Effect of Ethanol-exposure on Innate Immune Response in Sepsis.
Alcohol use disorder, reported by one in eight critically ill patients, is a risk factor for death in sepsis patients. Sepsis, the leading cause of death kills over 270,000 patients in the United States alone and remains without targeted therapy. Immune response in sepsis transitions from an early hyper-inflammation to persistent inflammation and immunosuppression and multiple organ dysfunction during late sepsis. Innate immunity is the first line of defense against pathogen invasion. Ethanol exposure is known to impair innate and adaptive immune response and bacterial clearance in sepsis patients. Specifically, ethanol-exposure is known to modulate every aspect of innate immune response with and without sepsis. Multiple molecular mechanisms are implicated in causing dysregulated immune response in ethanol-exposure with sepsis, but targeted treatments have remained elusive. In this manuscript, we outline the effects of ethanol-exposure on various innate immune cell types in general and during sepsis.


Review and analysis of strengths and weaknesses of agro-ecosystem models for simulating C and N fluxes.
Biogeochemical simulation models are important tools for describing and quantifying the contribution of agricultural systems to C sequestration and GHG source/sink status. The abundance of simulation tools developed over recent decades, however, creates a difficulty because predictions from different models show large variability. Discrepancies between the conclusions of different modelling studies are often ascribed to differences in the physical and biogeochemical processes incorporated in equations of C and N cycles and their interactions. Here we review the literature to determine the state-of-the-art in modelling agricultural (crop and grassland) systems. In order to carry out this study, we selected the range of biogeochemical models used by the CN-MIP consortium of FACCE-JPI (http://www.faccejpi.com): APSIM, CERES-EGC, DayCent, DNDC, DSSAT, EPIC, PaSim, RothC and STICS. In our analysis, these models were assessed for the quality and comprehensiveness of underlying processes related to pedo-climatic conditions and management practices, but also with respect to time and space of application, and for their accuracy in multiple contexts. Overall, it emerged that there is a possible impact of ill-defined pedo-climatic conditions in the unsatisfactory performance of the models (46.2%), followed by limitations in the algorithms simulating the effects of management practices (33.1%). The multiplicity of scales in both time and space is a fundamental feature, which explains the remaining weaknesses (i.e. 20.7%). Innovative aspects have been identified for future development of C and N models. They include the explicit representation of soil microbial biomass to drive soil organic matter turnover, the effect of N shortage on SOM decomposition, the improvements related to the production and consumption of gases and an adequate simulations of gas transport in soil. On these bases, the assessment of trends and gaps in the modelling approaches currently employed to represent biogeochemical cycles in crop and grassland systems appears an essential step for future research.


Molecular identification of Cryptosporidium parvum from avian hosts.
Cryptosporidium species are protozoan parasites that infect humans and a wide variety of animals. This study was aimed at identifying Cryptosporidium species and genotypes isolated from avian hosts. A total of 90 samples from 37 different species of birds were collected throughout a 3-month period from April 2008 to June 2008 in the National Zoo of Kuala Lumpur, Malaysia. Prior to molecular characterization, all samples were screened for Cryptosporidium using a modified Ziehl-Neelsen staining technique. Subsequently samples were analysed with nested-PCR targeting the partial SSU rRNA gene. Amplicons were sequenced in both directions and used for phylogenetic analysis using Neighbour-Joining and Maximum Parsimony methods. Although 9 (10%) samples were positive for Cryptosporidium via microscopy, 8 (8.9%) produced amplicons using nested PCR. Phylogenetic trees identified all the isolates as Cryptosporidium parvum. Although C. parvum has not been reported to cause infection in birds, and the role of birds in this study was postulated mainly as mechanical transporters, these present findings highlight the significant public health risk posed by birds that harbour the zoonotic species of Cryptosporidium.


Insulin-like growth factor-1 and its binding proteins during a low-protein diet in chronic renal failure.
The effects of a low-protein diet on the serum insulin-like growth factor (IGF)-1 and IGF binding proteins (IGFBP) were investigated during a 3-month controlled study in 12 adult chronic renal failure patients. Six patients were randomly supplemented with keto acids (Cetolog, Clintec, Velizy, France). Protein intake was prescribed so that both groups were isonitrogenous. Dietary survey included a monthly 3-day food record and a 24-h urinary urea measurement. After a 4- to 6-wk equilibrium period (1.11 g of protein, 32 kcal/kg body wt per day), patients reduced their protein intake to 0.71 g protein/kg body wt per day. Energy intake was kept constant (31 kcal/kg body wt per day) during the 3-month period. Serum IGF-1 levels were in normal range and, for 11 of the 12 patients, were correlated with the GFR (P = 0.01). These serum IGF-1 values did not decrease after reducing the protein intake. By Western ligand blotting, serum IGFBP1, IGFBP2, and IGFBP4 levels were significantly higher than normal adults, whereas the IGFBP3 level was not increased. IGFBP were not modified when protein intake was reduced. The IGFBP1 level was elevated despite a normal insulin level. IGFBP4 changes were inversely correlated with IGF-1 variations. There was no difference between groups receiving or not receiving the keto acids. Thus, in adult chronic renal failure, reducing protein intake by 40% did not modify the growth hormone/IGF-1/IGFBP axis.


Effect of 2.45 mT sinusoidal 50 Hz magnetic field on Saccharomyces cerevisiae strains deficient in DNA strand breaks repair.
To investigate whether extremely-low frequency magnetic field (MF) exposure produce alterations in the growth, cell cycle, survival and DNA damage of wild type (wt) and mutant yeast strains. wt and high affinity DNA binding factor 1 (hdf1), radiation sensitive 52 (rad52), rad52 hdf1 mutant Saccharomyces cerevisiae strains were exposed to 2.45 mT, sinusoidal 50 Hz MF for 96 h. MF was generated by a pair of Helmholtz coils. During this time the growth was monitored by measuring the optical density at 600 nm and cell cycle evolution were analysed by microscopic morphological analysis. Then, yeast survival was assayed by the drop test and DNA was extracted and electrophoresed. A significant increase in the growth was observed for rad52 strain (P = 0.005, Analysis of Variance [ANOVA]) and close to significance for rad52 hdf1 strain (P = 0.069, ANOVA). In addition, the surviving fraction values obtained for MF-exposed samples were in all cases less than for the controls, being the P value obtained for the whole set of MF-treated strains close to significance (P = 0.066, Student's t-test). In contrast, the cell cycle evolution and the DNA pattern obtained for wt and the mutant strains were not altered after exposure to MF. The data presented in the current report show that the applied MF (2.45 mT, sinusoidal 50 Hz, 96 h) induces alterations in the growth and survival of S. cerevisiae strains deficient in DNA strand breaks repair. In contrast, the MF treatment does not induce alterations in the cell cycle and does not cause DNA damage.